Rebecca A. Cole

BIOLOGICAL AND MOLECULAR CHARACTERIZATIONS OF TOXOPLASMA GONDII STRAINS OBTAINED FROM SOUTHERN SEA OTTERS (ENHYDRA LUTRIS NEREIS)

Toxoplasma gondii was isolated from brain or heart tissue from 15 southern sea otters (Enhydra lutris nereis) in cell cultures. These strains were used to infect mice that developed antibodies to T. gondii as detected in the modified direct agglutination test and had T. gondii tissue cysts in their brains at necropsy. Mouse brains containing tissue cysts from 4 of the strains were fed to 4 cats. Two of the cats excreted T. gondii oocysts in their feces that were infectious for mice. Molecular analyses of 13 strains indicated that they were all type II strains, but that they were genetically distinct from one another.

Vertical transmission of Neospora caninum in dogs

Herein we report the development of a canine model to examine transplacental and/or transmammary transmission of Neospora caninum. Six pregnant bitches were experimentally infected with N. caninum on day 21 of gestation. One bitch successfully delivered a litter of 3 live pups. These pups showed slight proprioception deficits, increased muscle tone, and spasticity in both pelvic limbs; however, N. caninum was neither demonstrated by bioassay in cell culture nor seen in histological sections of tissues from these pups. The remaining 5 bitches aborted. Neospora caninum was isolated from tissues of 4 of the experimentally infected bitches and pups from 2 of the miscarried litters. Experimental infections of 2 litters of 5-day-old nursing pups produced variable results. Neospora caninum was isolated from 2 of 2 inoculated pups in 1 litter and 3 of 6 inoculated pups in the second litter.

Relationship of tadpole stage to location of echinostome cercariae encystment and the consequences for tadpole survival.

The effect of echinostome infections on the survival of Rana pipiens tadpoles was examined in relation to developmental stage of tadpoles. Individual tadpoles of Gosner stages 25, 27, 32–33, and 37–39 were exposed to 1 of 4 levels of cercariae (0, 20, 50, or 100). Only tadpoles at stage 25, the earliest stage infected, died within a 5-day experimental period. This stage-specific mortality rate could be explained, in part, by the stage-specific location of encystment of cercariae, which was documented in a separate experiment. In accordance with kidney development, cercariae predominately encysted in the pronephroi during early stages of tadpole development (stages 25 through 31–32) and only in the mesonephroi and associated ducts at later stages (stages 37 through 46). As the mesonephros develops, renal capacity presumably increases. Thus, tadpoles died only when metacercariae concentrated in the functional portion of the kidney with the most limited renal capacity. As tadpoles aged, they also became less susceptible to infections. On average, 69.5% of cercariae that were exposed to stage 25–26 tadpoles successfully encysted, compared with only 8.4% of cercariae exposed to stage 37–38 tadpoles. Exposures of metamorphic frogs (poststage 46) to cercariae revealed that these individuals can become infected with echinostomes. Collectively, our data highlight the host stage–dependent dynamics of tadpole–echinostome interactions.

Using occupancy models to understand the distribution of an amphibian pathogen, Batrachochytrium dendrobatidis

Batrachochytrium dendrobatidis is a fungal pathogen that is receiving attention around the world for its role in amphibian declines. Study of its occurrence patterns is hampered by false negatives: the failure to detect the pathogen when it is present. Occupancy models are a useful but currently underutilized tool for analyzing detection data when the probability of detecting a species is <1. We use occupancy models to evaluate hypotheses concerning the occurrence and prevalence of B. dendrobatidis and discuss how this application differs from a conventional occupancy approach. We found that the probability of detecting the pathogen, conditional on presence of the pathogen in the anuran population, was related to amphibian development stage, day of the year, elevation, and human activities. Batrachochytrium dendrobatidis was found throughout our study area but was only estimated to occur in 53.4% of 78 populations of native amphibians and 66.4% of 40 populations of nonnative Rana catesbeiana tested. We found little evidence to support any spatial hypotheses concerning the probability that the pathogen occurs in a population, but did find evidence of some taxonomic variation. We discuss the interpretation of occupancy model parameters, when, unlike a conventional occupancy application, the number of potential samples or observations is finite.

Helminths of the raccoon (Procyon lotor) in western Kentucky.

Seventy raccoons (Procyon lotor) from western Kentucky were examined for helminths from December 1985 through May 1986. Twenty-three species of helminths were collected including 10 species of Trematoda (Brachylaima virginiana, Euryhelmis squamula, Eurytrema procyonis, Fibricola cratera, Gyrosoma singulare, Maritreminoides nettae, Mesostephanus appendiculatoides, Metagonimoides oregonensis, Paragonimus kellicotti, Pharyngostomoides procyonis), 2 species of Cestoda (Atriotaenia procyonis, Mesocestoides variabilis), 10 species of Nematoda (Arthrocephalus lotoris, Baylisascaris procyonis, Capillaria putorii, C. plica, Crenosoma goblei, Dracunculus insignis, Gnathostoma procyonis, Molineus barbatus, Physaloptera rara, Trichinella spiralis), and 1 species of Acanthocephala (Macracanthorhynchus ingens). A mean of 6.4 (3-11) helminth species per host was recorded. Fibricola cratera, Atriotaenia procyonis, Mesocestoides variabilis, Arthrocephalus lotoris, Capillaria plica, Dracunculus insignis, Molineus barbatus, and Physaloptera rara were ubiquitous parasites of the raccoon, whereas specific nidi were observed for Eurytrema procyonis, Gyrosoma singulare, Paragonimus kellicotti, Baylisascaris procyonis, Trichinella spiralis, and Macracanthorhyncus ingens. With an overall prevalence of 10% or higher, 15 of the 23 helminth species were considered common parasites of the raccoon in western Kentucky. When the 10% prevalence rate was applied within geographical quadrants to correct for the presence of nidi it was found that 18 of the 23 helminth species were common and 5 were regarded as rare parasites of the raccoon. Two species of nematodes, T. spiralis and B. procyonis, displayed a markedly higher prevalence in male raccoons.

Effects of wetland vs. landscape variables on parasite communities of Rana pipiens: links to anthropogenic factors

The emergence of several diseases affecting amphibian populations worldwide has prompted investigations into determinants of the occurrence and abundance of parasites in frogs. To understand the spatial scales and identify specific environmental factors that determine risks of parasitism in frogs, helminth communities in metamorphic frogs of the northern leopard frog (Rana pipiens) were examined in relation to wetland and landscape factors at local (1 km) and regional (10 km) spatial extents in an agricultural region of Minnesota (USA) using regression analyses, ordination, and variance partitioning techniques. Greater amounts of forested and woody wetland habitats, shorter distances between woody wetlands, and smaller-sized open water patches in surrounding landscapes were the most consistently positive correlates with the abundances, richness, and diversity of helminths found in the frogs. Wetland and local landscape variables were suggested as most important for larval trematode abundances, whereas local and regional landscape variables appeared most important for adult helminths. As previously reported, the sum concentration of atrazine and its metabolite desethylatrazine, was the strongest predictor of larval trematode communities. In this report, we highlight the additional influences of landscape factors. In particular, our data suggest that anthropogenic activities that have resulted in the loss of the availability and connectivity of suitable habitats in the surrounding landscapes of wetlands are associated with declines in helminth richness and abundance, but that alteration of wetland water quality through eutrophication or pesticide contamination may facilitate the transmission of certain parasite taxa when they are present at wetlands. Although additional research is needed to quantify the negative effects of parasitism on frog populations, efforts to reduce inputs of agrochemicals into wetlands to limit larval trematode infections may be warranted, given the current high rates of amphibian declines and extinction events.

Parasites of native and nonnative fishes of the Little Colorado River, Grand Canyon, Arizona.

A 2-yr, seasonal, parasitological study of 1,435 fish, belonging to 4 species of native fishes and 7 species of nonnative fishes from the lower Little Colorado River (LCR) and tributary creeks, Grand Canyon, Arizona, yielded 17 species of parasites. These comprised 1 myxozoan (Henneguya exilis), 2 copepods (Ergasilus arthrosis and Lernaea cyprinacea), 1 acarine (Oribatida gen. sp.), 1 piscicolid leech (Myzobdella lugubris), 4 monogeneans (Gyrodactylus hoffmani, Gyrodactylus sp., Dactylogyrus extensus, and Ligictaluridus floridanus), 4 nematodes (Contracaecum sp., Eustrongylides sp., Rhabdochona sp., and Truttaedacnitis truttae), 3 cestodes (Bothriocephalus acheilognathi, Corallobothrium fimbriatum, and Megathylacoides giganteum), and 2 trematodes (Ornithodiplostomum sp. and Posthodiplostomum sp.). Rhabdochona sp. was the only adult parasite native to the LCR. Infection intensities of Ornithodiplostomum sp. and B. acheilognathi were positively correlated with length of the humpback chub Gila cypha. Adult helminths showed a high degree of host specificity, except B. acheilognathi, which was recovered from all fish species examined but was most abundant in cyprinids. Abundance of B. acheilognathi in the humpback chub was highest in the fall and lowest in the summer in both reaches of the LCR. There was no major taxonomic difference in parasite assemblages between the 2 different reaches of the river (LC1 and LC2). Parasite community diversity was very similar in humpback chub, regardless of sampling site or time. The parasite fauna of the LCR is numerically dominated by B. acheilognathi and metacercariae of Ornithodiplostomum sp. The richest and most diverse component community occurred in a nonnative species, the channel catfish Ictalurus punctatus, but infracommunity species richness was highest in a native host, humpback chub.

Experimental infection of the endangered bonytail chub (Gila elegans) with the Asian fish tapeworm (Bothriocephalus acheilognathi): impacts on survival, growth, and condition

Bothriocephalus acheilognathiYamaguti, 1934, a tapeworm known to be pathogenic to some fish species, has become established in the endangered humpback chub (Gila cypha Miller, 1964) in Grand Canyon, USA, following the tapeworms introduction into the Colorado River system. The potential impact of this tapeworm on humpback chub was studied by exposing the closely related bonytail chub (Gila elegans Baird and Girard, 1853) to the parasite under a range of conditions that included potential stressors of humpback chub in their natal waters, such as abrupt temperature change and a limited food base. Survival of infected fish under low food rations was considerably lower than that of control fish, and mortality of infected fish began 20 days earlier. Growth of infected fish was signifi- cantly reduced, and negative changes in health condition indices were found. No significant negative impacts were re- vealed from the synergistic effects between temperature shock and infection. Bothriocephalus acheilognathi does present a potential threat to humpback chub in Grand Canyon and should be considered, along with conventional con- cerns involving altered flow regimes and predation, when management decisions are made concerning conservation of this endangered species. Resume ´ : Bothriocephalus acheilognathiYamaguti, 1934, un ver plat connu comme pathogene pour certaines especes de

Epizootic podoknemidokoptiasis in American robins.

Epizootics of scaly leg disease caused by infection with the submacroscopic mite Knemidokoptes jamaicensis (Acari: Knemidokoptidae) in migratory American robins (Turdus migratorius) from a residential area of Tulsa (Oklahoma, USA) are documented during the winters (December through February) of 1993–94 and 1994–95. Estimates of 60 to >80% of the birds in several different flights arriving in the area had lesions consistent with knemidokoptic mange. Epizootic occurrence of K. jamaicensis also is confirmed incidentally in American robins from Georgia (USA) in 1995 and 1998 and in Florida (USA) in 1991. These are the first confirmed epizootics of scaly leg attributed to infections with mites specifically identified as K. jamaicensis in North America. Severity of observed lesions in American robins ranged from scaly hyperkeratosis of the feet and legs to extensive proliferative lesions with loss of digits or the entire foot in some birds. Histologically, there was severe diffuse hyperkeratosis of the epidermis which contained numerous mites and multifocal aggregates of degranulating to degenerating eosinophilic heterophils; there was mild to severe superficial dermatitis with aggregates of eosinophilic heterophils and some mononuclear cells. Based on limited data from affected captive birds in Florida, we questioned the efficacy of ivermectin as an effective acaricide for knemidokoptiasis and propose that conditions associated with captivity may exacerbate transmission of this mite among caged birds. While knemidokoptic mange apparently can result in substantial host morbidity and possibly mortality, the ultimate impact of these epizootics on American robin populations presently is unknown.

PREVALENCE OF SKELETAL AND EYE MALFORMATIONS IN FROGS FROM NORTH-CENTRAL UNITED STATES: ESTIMATIONS BASED ON COLLECTIONS FROM RANDOMLY SELECTED SITES

Skeletal malformation rates for several frog species were determined in a set of randomly selected wetlands in the north-central USA over three consecutive years. In 1998, 62 sites yielded 389 metamorphic frogs, nine (2.3%) of which had skeletal or eye malformations. A subset of the original sites was surveyed in the following 2 yr. In 1999, 1,085 metamorphic frogs were collected from 36 sites and 17 (1.6%) had skeletal or eye malformations, while in 2000, examination of 1,131 metamorphs yielded 16 (1.4%) with skeletal or eye malformations. Hindlimb malformations predominated in all three years, but other abnormalities, involving forelimb, eye, and pelvis were also found. Northern leopard frogs (Rana pipiens) constituted the majority of collected metamorphs as well as most of the malformed specimens. However, malformations were also noted in mink frogs (R. septentrionalis), wood frogs (R. sylvatica), and gray tree frogs (Hyla spp.). The malformed specimens were found in clustered sites in all three years but the cluster locations were not the same in any year. The malformation rates reported here are higher than the 0.3% rate determined for metamorphic frogs collected from similar sites in Minnesota in the 1960s, and thus, appear to represent an elevation of an earlier baseline malformation rate.